Functional supersensitivity of antinociceptive spinal cord alpha 2-adrenoceptors, induced by depletion of endogenous noradrenaline, is associated with an enhanced sensitivity for guanine nucleotide regulation of 3H-clonidine binding.

In spinal cords from normal mice, 3H-clonidine bound to alpha 2-adrenoceptors with an apparant Kd of 4.6 nM and capacity (Bmax) of 430 fmol/mg protein. The non-hydrolyzable GTP analogue Gpp(NH)p was found to dose-dependently reduce the binding of 3H-clonidine the effect of Gpp(NH)p being essentially maximal at 10(-4) M. Gpp(NH)p was found to reduce the apparent affinity as well as the apparent number of binding sites for 3H-clonidine; the Kd of 3H-clonidine being 16 nM and the Bmax 300 fmol/mg protein when 10(-4) M of the nucleotide was present. In mice pretreated with the noradrenaline neurotoxin DSP4 for 14 days prior to assay, the specific binding of nearly saturating concentrations (10-30 nM) of 3H-clonidine was virtually the same as for control mice. However, for the DSP4 treated animals the down-regulation of 3H-clonidine binding induced by 10(-4) M Gpp(NH)p was significantly higher than for control mice. Thus, at a 3H-clonidine concentration of 10 nM the down-regulation induced by Gpp(NH)p was 64.1 +/- 2.6% for the DSP4 versus 58.9 +/- 2.3% for the control mice (P less than 0.05). At a 3H-clonidine concentration of 30 nM the down-regulation was 52.4 +/- 1.4% for DSP4 versus 41.8 +/- 4.1% for control mice (P less than 0.05). DSP4 pretreatment also potentiated the antinociceptive effect of intrathecal clonidine, as assessed by the tail-flick and hot plate tests. It is suggested that the functional supersensitivity induced by DPS4 for clonidine is related to the increased sensitivity for guanine nucleotide regulation of the spinal cord alpha 2-adrenoceptors.